, Volume 18, Issue 8, pp 437–442 | Cite as

Mycorrhizal morphotyping and molecular characterization of Chondrogaster angustisporus Giachini, Castellano, Trappe & Oliveira, an ectomycorrhizal fungus from Eucalyptus

  • M. Lupatini
  • P. A. P. Bonnassis
  • R. B. Steffen
  • V. L. Oliveira
  • Z. I. Antoniolli
Short Note


Chondrogaster angustisporus is a hypogeous ectomycorrhizal fungus described from fruiting bodies collected under Eucalyptus spp. in Brazil, Uruguay, and Australia. Due to its efficiency in promoting plant growth, we decided to characterize this fungus through mycorrhizal morphotyping and internal transcribed spacer (ITS) (rRNA) sequencing. DNA extracted from mycelium was amplified and sequenced using specific primers. Mycorrhizas were obtained aseptically and analyzed in terms of macroscopic and microscopic characteristics. When compared with other fungal DNA sequences available in the NBCI GenBank, the C. angustisporus sequence presented the highest similarity to an uncultured ectomycorrhizal fungus from the Seychelles. It also shows significant similarities to Gomphus, Ramaria, and Hysterangium species supporting the classification of Chondrogaster in the subclass Phallomycetidae in the gomphoid–phalloid group. The mycorrhizas were characterized by a narrow mantle with a single tissue layer densely arranged and organized as a net synenchyma with elongated hyphae. Interhyphal spaces were seen only in the external region where hyphae were more loosely organized. Bottle-shaped cystidia with bent necks were observed on the surface of the mantle. Emanating hyphae were larger than those in the mantle and presented a granular content. At regular intervals the hyphae were divided by septa with clamp connections. The Hartig net was of the common type, with typical palmetti and single hyphal rows and limited to the epidermal layer. The mycorrhizal description and the ITS sequence obtained are useful tools to identify this ectomycorrhizal fungus in culture and in association with Eucalyptus roots.


Mycorrhizal morphotyping Internal transcribed spacer (ITS) Sequencing Molecular identification 


  1. Agerer R (1987–1993) Colour atlas of ectomycorrhizae. Einhorn-Verlag Eduard Dietenberger, Schwäbisch GmündGoogle Scholar
  2. Altschul SF, Madden TL, Schaffon AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein data base search programs. Nucleic Acids Research 25:3389–3402PubMedCrossRefGoogle Scholar
  3. Alvarez ML, Cerceda MC (2005) Chondrogaster angustisporus, nueva cita para Europa. Cryptogam, Mycol 26:113–122Google Scholar
  4. Bougher NL, Castellano MA (1993) Delimitation of Hymenogaster sensu stricto and four new segregate genera. Mycologia 85:273–293 doi:10.2307/3760462 CrossRefGoogle Scholar
  5. Bougher NL, Lebel T (2001) Sequestrate (truffle-like) fungi of Australia and New Zealand. Aust Syst Bot 14:439–484 doi:10.1071/SB00002 CrossRefGoogle Scholar
  6. Bougher NL, Syme K (1998) Fungi of Southern Australia. University of Western Australia Press, NedlandsGoogle Scholar
  7. Brundrett M, Bougher NL, Dell B, Grove T, Malajczuk N (1996) Working with mycorrhizas in forestry and agriculture. ACIAR Monograph 32, CanberraGoogle Scholar
  8. Castellano MA, Bougher NL (1994) Consideration of the taxonomy and biodiversity of Australian ectomycorrhizal fungi. Plant Soil 159:37–46Google Scholar
  9. Castellano MA, Trappe JM, Maser Z, Maser C (1989) Key to spores and the genera of hypogeous fungi of North temperate forests, with special reference to animal mycophagy. Mad River, EurekaGoogle Scholar
  10. Dunn IS, Blattner FR (1987) Charon-36 to charon-40 multi enzyme, high-capacity, recombination deficient replacement vectors with polylinkers and polystuffers. Nucleic Acids Res 15:2677–2698 doi:10.1093/nar/15.6.2677 PubMedCrossRefGoogle Scholar
  11. Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes—application to the identification of mycorrhizae and rusts. Mol Ecol 2:113–118 doi:10.1111/j.1365-294X.1993.tb00005.x PubMedCrossRefGoogle Scholar
  12. Giachini AJ, Oliveira VL, Castellano MA, Trappe JM (2000) Ectomycorrhizal fungi in Eucalyptus and Pinus plantations in southern Brazil. Mycologia 92:1166–1177 doi:10.2307/3761484 CrossRefGoogle Scholar
  13. Giachini AJ, Souza LAB, Oliveira VL (2004) Species richness and seasonal abundance of ectomycorrhizal fungi in plantations of Eucalyptus dunnii and Pinus taeda in southern Brazil. Mycorrhiza 14:375–381PubMedGoogle Scholar
  14. Goodman DM, Durall DM, Trofymow JA, Berch SM (1996–1998) A manual of concise descriptions of North American ectomycorrhizae. Mycologue Publications, SydneyGoogle Scholar
  15. Hosaka K, Bates ST, Beever RE, Castellano MA, Colgan W, Dominguez LS et al (2006) Molecular phylogenetics of the gomphoid–phalloid fungi with an establishment of the new subclass Phallomycetidae and two new orders. Mycologia 98:949–959 doi:10.3852/mycologia.98.6.949 PubMedCrossRefGoogle Scholar
  16. Ingleby K, Mason PA, Last FT, Fleming LV (1990) Identification of ectomycorrhizas. Institute for Terrestrial Ecology, Natural Environmental Research Council, U.K. Res. Publ. No. 5, LondonGoogle Scholar
  17. Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163 doi:10.1093/bib/5.2.150 PubMedCrossRefGoogle Scholar
  18. LASERGENE (1994) User’s guide: a manual for the Lasergene System. Madison. Biocomputing software for Windows, 253 pGoogle Scholar
  19. Lima WP (1993) Impacto Ambiental do Eucalipto, 2nd edn. EDUSP, São PauloGoogle Scholar
  20. Lu X, Malajczuk N, Brundett M, Dell B (1999) Fruiting of putative ectomycorrhizal fungi under blue gum (Eucalyptus globulus) plantations of different ages in Western Australia. Mycorrhiza 8:255–261 doi:10.1007/s005720050242 CrossRefGoogle Scholar
  21. Marx DH (1969) The influence of ectotrophic mycorrhizal fungi on the resistance of pine roots to pathogenic fungi and soil bacteria. Phytopatology 59:153–163Google Scholar
  22. Midgley DJ, Saleeba JA, Stewart MI, Simpson AE, McGee PA (2007) Molecular diversity of soil basidiomycete communities in northern-central New South Wales, Australia. Mycol Res 111:370–378 doi:10.1016/j.mycres.2007.01.011 PubMedCrossRefGoogle Scholar
  23. Norkrans B (1949) Some mycorrhiza-forming Tricholoma species. Sven Bot Tidskr 43:485–490Google Scholar
  24. Peterson RL, Massicotte HB, Melville LH (2004) Mycorrhizas: anatomy and cell biology. NRC Research Press, OttawaGoogle Scholar
  25. Rossi MJ (2006) Technology to the production of ectomycorrhizal fungi inoculants employing submerged cultivation in an airlift bioreactor, Ph.D. Thesis, Universidade Federal de Santa Catarina, Florianópolis, Brazil (in Portuguese) (
  26. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning—a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  27. Souza LAB, Silva Filho GN, Oliveira VL (2004) Efficiency of ectomycorrhizal fungi on phosphorus uptake and growth promoting of eucalyptus. Pesquisa Agropecu Bras 39:349–355 in PortugueseGoogle Scholar
  28. Tedersoo L, Suvi T, Beaver K, Koljalg U (2007) Ectomycorrhizal fungi of the Seychelles: diversity patterns and host shifts from the native Vateriopsis seychellarum (Dipterocarpaceae) and Intsia bijuga (Caesalpiniaceae) to the introduced Eucalyptus robusta (Myrtaceae), but not Pinus caribea (Pinaceae). New Phytol 175:321–333 doi:10.1111/j.1469-8137.2007.02104.x PubMedCrossRefGoogle Scholar
  29. White T, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols—a guide to methods and applications. Academic, New York, pp 315–322Google Scholar
  30. Yang CS, Wilcox HE (1983) Technique for observation of mycorrhizal development under monoxenic conditions. Can J Bot 62:251–254 doi:10.1139/b84-041 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • M. Lupatini
    • 1
  • P. A. P. Bonnassis
    • 2
  • R. B. Steffen
    • 1
  • V. L. Oliveira
    • 2
  • Z. I. Antoniolli
    • 1
  1. 1.Dept. de Solos, Centro de Ciências RuraisUniversidade Federal de Santa MariaSanta MariaBrazil
  2. 2.Dept. de Microbiologia e Parasitologia, Centro de Ciências BiológicasUniversidade Federal de Santa CatarinaFlorianópolisBrazil

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